GRB2 antibody

Catalog No. ABIN686932

This independently validated Rabbit Polyclonal antibody specifically detects GRB2 in ELISA and IHC (p). It exhibits reactivity toward Human, Mouse, Rat, Dog and Chicken.

Quick Overview for GRB2 antibody (ABIN686932)

Target: GRB2 (Growth Factor Receptor-Bound Protein 2 (GRB2))

Reactivity: Human, Mouse, Rat, Dog, Chicken

Host: Rabbit

Clonality: Polyclonal

Conjugate: This GRB2 antibody is un-conjugated

Application: ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Product Details anti-GRB2 Antibody

Cross-Reactivity: Human, Rat

Predicted Reactivity: Mouse,Dog,Chicken

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human GRB2

Isotype: IgG

Application Details

Application Notes: WB(1:100-500)
Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029728 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029728

Date: 06/05/2014

Lot Number: 121228

Method validated: Western Blotting

Positive Control: NIH/3T3 cells

Negative Control: HeLa cells - very low expression

Notes: A strong band was seen at the correct molecular weight in the positive control, and no band was present in the negative control. Additionally, no non-specific bands were observed.

Validation Images

Figure 1: Western blot of lysates from NIH/3T3 cells (Lane 1, positive control) and HeLa cells (Lane 2, negative control) probed with anti-GRB2 (upper panel) or with anti- Actin for loading control (lower panel).

Full Methods

Primary Antibody:

• Antigen: Growth Factor Receptor-Bound Protein 2 (GRB2) (1:200 dilution)
• Catalog number: ABIN686932
• Lot number: 121228

Secondary Antibody:

• Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
• Lot number: L170-6515

Full Protocol:

• Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
• 46 μg of boiled extracts were loaded and resolved on a 8-16% SDS-polyacrylamide gel.
• The Spectra Multicolor Broad Range (Thermo Scientific, Cat # 26634) were used as molecular mass markers.
• Proteins were transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) non-fat dry milk at room temperature for 1 h.
• The membrane was rinsed with TBST once.
• The membrane was immersed with the protein side up in the primary antibody solution (anti-GRB2; 1:200) in TBST containing 5% (W/V) non-fat dry milk and incubated for 16 hours at 4°C.
• The membrane was rinsed in TBST three times for 5 min each.
• The membrane was incubated in the HRP-conjugated secondary antibody solution (goat-anti rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) non-fat dry milk and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
• The membrane was rinsed three times with TBST for 5 min each.
• The membrane was rinsed in TBS twice for 30 s each.
• Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
• The membrane was rinsed three times with TBST, then incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
• The membrane was washed in TBST 2 times for 10 min each.
• Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (goat-anti rabbit IgG-HRP; 1:20,000).

Experimental Notes: Nothing to note.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

Target Details for GRB2

Target: GRB2 (Growth Factor Receptor-Bound Protein 2 (GRB2))

Alternative Name: GRB2/ASH

Background: Optional[synonyms]: GRB 2, GRB2 adapter protein, Grb3 3, Growth Factor Receptor Bound Protein 2, Growth factor receptor bound protein 3, EGFRBP GRB2, Growth factor receptor-bound protein 2, Abundant SRC homology, Adapter protein GRB2, ASH, Ash protein, EGFRBP GRB2, Epidermal growth factor receptor binding protein, Epidermal growth factor receptor binding protein GRB2, GRB 2, GRB2 adapter protein, Grb3 3, Growth Factor Receptor Bound Protein 2, Growth factor receptor bound protein 3, HT027, MST084, MSTP084, OTTHUMP00000166096, OTTHUMP00000166097, OTTHUMP00000166098, Protein ASH, SEM5, SH2/SH3 adapter GRB2.

Molecular Weight: 25kDa

Gene ID: 2885

UniProt: P62993

Pathways: RTK Signaling, TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Actin Filament Polymerization, Hepatitis C, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, EGFR Downregulation